C-12-substituted progesterone



United States 1 3,047,570 C-12-SUBSTITUTED PROGESTERONE Otto Halpern, Mexico City, Mexico, assignor, by mesne assignments, to Syntex Corporation, a corporation of Panama No Drawing. Filed Feb. 10, 1961, Ser. No. 88,259 9 Claims. (Cl. 260-23957) COOR I CH3 A Ow pt In the above formulas R and R represent hydrogen or a lower alkyl group and R represents a hydrogen or an acyl radical containing up to 12 carbon atoms.

The novel compounds of the present invention are prepared by a method illustrated by the following equation:

w 1 an,

atent 3,047,570 Patented July 31, 1962 In the above formulas R represents hydrogen or a lower alkyl group and Ac represents the acyl radical of a hydrocarbon carboxylic acid containing from 1 to 12 carbon atoms.

The wavy line at C-12 indicates a generic expression for the a and B steric configurations.

In practicing the process outlined above, botogenin acetate (I) is refluxed in a solvent such as anhydrous benzene for 2 to 3 hours with zinc and an ester of an orhalo carboxylic acid such as for example a-bromoaeetic acid. The reaction product is washed with acid and then saponified as by boiling with dilute methanolic potassium carbonate for 6-8 hours. The crude acidic material (II) is then esterified as by treatment with excess ethereal diazoalkane or by the Fischer method, then acylated, preferably acetylated with acetic anhydride in pyridine solution on the steam bath for one hour to form the C-12 isomeric hydroxy esters represented by Formula III. Degradation of the spiroketal side chain is then effected by conventional procedure as by reaction with acetic anhydride at about 200, oxidation of the resulting pseudo compound to the diosone followed by alkaline hydrolysis, esterification with a diazoalkane and acylation in pyridine solution to thus form 1Z-carboalkoxymethylene-A pregnadien-Bfi-ol-ZO-one acylate (IV). The C-2O keto group is then protected by reaction with ethylene glycol in the presence of p-toluenesulfonic acid to form the cyclic ethylene ketal (VII). Selective reduction of the C-12 exocyclic bond is then effected with concomitant reduction of the carboalkoxy group at C-l2 and hydrolysis of the ester group at C-3 by reaction with lithium in liquid ammonia, thus producing the 20-cyclic ethylene ketal of 12fl-hydroxyethyl-A -pregnadien-3B-ol-20-one (VIII). The ketal group is hydrolyzed by reaction with perchloric acid in tetrahydrofuran and there is regenerated the 0-20 keto group thus affording 12fl-hydroxyethyl-A -pregnadien-3fi-ol-20-one.

For introduction of a hydroxyl group at C-l7oz, the 16,17-double bond is first epoxidized, preferably by reaction with aqueous alkaline peroxide; the resulting 16,17- epoxide is reacted with hydrogen bromide to form the 16B-bromo-17a-hydroxy grouping which upon treatment with hydrogen in a solvent such :as methanol and in the presence of ammonium acetate and a palladium catalyst results in reductive debromination and there is formed 12,8 hydroxyethyl A pregnene 35,170: diol 20- one (IX). Upon subjecting the latter compound to Jones oxidation, the Bfl-hydroxy group is oxidized to the keto group and the 12-hydroxyethyl group is oxidized to the acid. The resulting B-keto 12-acetic acid compound is treated with a strong mineral acid, such as hydrochloric acid in acetic acid solution to effect formation of the lactone ring with concomitant shifting of the double bond to C-4,5 thus producing the C-1 2,l' 7a-l actone of l2=carboxymethyl-17u-hydroxy-progesterone [C-12,17a-lactone of A -pregnen-17 t-ol-3,20-dione-12-acetic acid (X)] Alternatively, the 3-keto compound obtained after Jones oxidation is treated with oxalic acid in methanol solution, to produce the 0:,[3 unsaturated ketone, namely 1ZB-carboxymethyl-A -pregnen-17fi-ol-3,20 dione, and this compound treated with a strong mineral acid, acid ansten-3/8-ol (XIII).

In the above equation R has the same meaning as previously set forth.

In practicing the process outlined above, the C-12 isomeric hydroxy esters (-III) are heated with acetic anhydried in a sealed tube at about 200 C. to form the carboxymethylene derivative (XI). The latter is then treated with dilute methanolic potassium hydroxide solution to afford .the l2-carboxymethylene derivative of A -22a-spirosten- 318-01 (XII) which upon reduction under Birch conditions is converted into l2,8-carboxymethyl-A -22a-spiro- Degradation of the spiroketal side chain as described previously affords IZB-carboxymethyl- A pregnadien-3,B-ol--one (XIV) which upon esterification with a diazoalkane affords the l-carboa'lkoxymethyl-A -pregnadien-3 3-ol-20-one derivative. droxy group is then introduced at C-17oc in the same manner described above to form the IZB-carboxymethyl or 12 8-carboalkoxymethyl-A -pregnen-3 8,l7u-diol 20 one (XV). Upon oxidation with chromic acid there is formed the IZB-oarboxymethyl or lZB-carboalkoxymethyl-A pregnen-17a-ol-3,20-dione which upon treatment with acid as set forth previously is transformed into the 12/8,

- 17oz lactone of A -pregnen17u.-o1-3,ZO-dione-12-acetic acid (X).

Other (ac-halogenated esters such as, for example, the

4 esters of a-chloro or a-iodo propionic acid, a-bromobutyric acid, a-bromocaproic acid are employed instead of a-brom0acetic acid to form the corresponding 12-carboxylic acid derivative.

The following examples serve to illustrate but are not intended to limit the scope of the present invention:

Example I To a solution of 50 g. of botogenin acetate in 500 cc. of absolute benzene were added 75 cc. of bromoacetic acid and 75 g. of iodine-activated zinc granules. The mixture was refluxed for 3 hours, evaporated and taken up in ether. The ether layer was washed with 2 N HCl, water, sodium bicarbonate solution and finally with water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness; the residue was dissolved in 700 cc. of methanol, treated with 150 g. of potassium carbonate previously dissolved in 300 cc. of water and the reaction mixture was refluxed for 8 hours, the methanol was then evaporated under vacuo and the aqueous suspension diluted with water, extracted with ether several times, and acidified, to precipitate a mixture of the 12oz and 12B- hydroxy acids, that was collected by filtration.

The above crude acidic material was refluxed for 3 hours with 100 cc. of methanol containing 3.6 cc. of sulfuric acid, diluted with water, the methanol evaporated under vacuo, and the product extracted several times with ether, the organic extract was washed then with 5% sodium bicarbonate solution and water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness. The residue was dissolved in 50 cc. of pyridine and 50 cc. of acetic anhydride and the mixture heated for 1 hour on the steam bath, poured into ice water and stirred for 30 minutes to hydrolyze the excess of anhydride. It was then extracted with ethyl acetate, the organic extract washed well with water, dried and evaporated to dryness, the residue consisting of a mixture of the l2-isomeric hydroxy esters, namely l2a-carb-0methoxymethyl-A -22aspirosten-3fi,l2/3-d-iol-3 acetate and l2fi-carbomethoxymethyl-A -22a-spirosten-3fi,12a-dio1 3-acetate, was purified by filtration on Washed alumina in benzene solution.

Example I] 10 g. of the above mixture of hydroxyesters was heated for 5 hours in a sealed tube, at 195 C., with 40 cc. of acetic anhydride. The contents of the tube were then transferred to an Erlenmeyer, 10 cc. of water was added cautiously, and heated for half an hour on the steam bath to decompose the excess of anhydride. The solution was cooled to about 15 C. and treated under stirring with 50 cc. of a precooled solution of chromium trioxide in acetic acid (prepared by dissolving 4.2 g. of chromium trioxide in 54.5 cc. of 90% acetic acid). The reaction mixture was stirred for 30 minutes further, the excess of reagent was destroyed with sodium bisulfite solution and the mixture poured into water, extracted with ethyl acetate, the extracts washed with water, sodium bicarbonate solution and Water to neutral, dried and evaporated to dryness. The residue was refluxed for 1 hour with a mixture of 250 cc. of t-butanol and cc. of 20% aqueous potassium hydroxide, it was then poured into water, extracted well with ethyl acetate, the aqueous layer acidified and reextracted also with ethyl acetate. The acid extract was dried and evaporated to dryness thus giving the crude 12-carboxymethylene-A -pregnadien-35-ol-20-one.

Esterification of this compound with an excess of diazomethane or by the Fischer method, in accordance with the procedure of Example I, gave l2-carbomethoxymethylene-A -pregnadien-3fl-ol-ZO-one.

500 mg. of the crude ester was acetylated with 2 cc. of pyridine and 1 cc. of acetic anhydride in the usual manner (1 hour on the steam bath). Chromatography of the crude reaction product on neutral alumina gave the acetate of l2-carbomethoxymethylene-A -pregnadien- 3,B-ol-20-one, M.P. 169-7 1 C.; A max 228 m log. 4.28.

5 This compound exhibited anti-androgenic, anti-estrogenic and anti-ovulatory activity as well as gonadotrophin suppression.

Example III A suspension of 500 mg. of the above compound in 60 cc. of ethylene glycol was treated with 40 mg. of ptoluenesulfonic acid and the mixture heated in an oil bath to approximately 90 C. under high vacuum. After 5 hours the homogeneous solution was concentrated to approximately 10 cc. by heating to about 120 C. under high vacuum. The concentrate was poured into dilute aqueous sodium bicarbonate solution, extracted with ethyl acetate and the organic extract washed to neutrality, dried over anhydrous sodium sulfate and evaporated to dryness, thus producing ZO-ethylenedioxy-ll-carbornethoxymethylene-A -pregnadien-3fl-o1-3-acetate that was not further purified.

The above crude compound was dissolved in 4 cc. of anhydrous dioxane and 4 cc. of anhydrous ether and added dropwise to a solution of lithium in liquid ammonia in such a way that the solution remains blue throughout the addition; after stirring another minute or two, the blue color was discharged by adding solid ammonium chloride. The ammonia was then evaporated and the residue taken up in water and extracted with ethyl acetate. After washing with dilute hydrochloric acid and Water to neutral, the organic extract was dried and evaporated under reduced pressure, thus giving the 20-ethylene ketal of 1ZB-hydroxyethyl-M -pregnadien- 3B-ol-20-one.

The above compound was disolved in 10 cc. of tetrahydrofuran and treated with 4 cc. of 3 N perchloric acid, the reaction mixture was kept at room temperature for 3 hours, poured into water, extracted with methylene chloride and the organic extract washed with water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness. Chromatography of the residue afforded the pure 1Zfi-hydroxyethyl-A -pregnadien-3B-ol-20-one.

Example IV A solution of 0.4 g. of the latter compound in 16 cc. of methanol was treated at C. with 1 cc. of 35% hydrogen peroxide and 0.4 g. of potassium hydroxide previously dissolved in 1.6 cc. of water and the mixture kept at 0 C. overnight, diluted with ice-salt water, and extracted several times with ether. The combined extracts were washed with water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness. Crystallization from acetone-hexane gave 12,B-hydroxyethyl-16a,17a-oxido-A pregnen-3B-ol-20-one.

The preceding 16,17-epoxide, dissolved in cc. of acetic acid, was treated at room temperature and while stirring, with 0.1 cc. of a saturated solution of hydrobromic acid in glacial acetic acid. After half an hour it was poured into water, the precipitate was filtered and washed with water to neutrality. Thus there was obtained 16/3-bromo-lZfl-hydroxyethyl A pregnene-3p,l7a-diol- 20-one.

The above bromohydrin was dissolved in cc. of methanol and stirred overnight with 300 mg. of 2% palladium on calcium carbonate and 10 mg. of ammonium acetate under an atmosphere of hydrogen. The suspension was filtered through celite, the filter was washed with hot methanol, the washings and filtrate were combined and then evaporated to dryness. By crystallization from methanol there was obtained 12fi-hydroxyethyl-A pregnene-3,8,17zx-diol--one which exhibited anti-androgenic, anti-estrogenic and progestational activity as well as suppression of the gonadotrophic hormone.

Example V 1 g. of 1Zfl-hydroxyethyl-A -pregnene-3[5,17a-diol-20- one was dissolved in 50 cc. of acetone, cooled to 0 C., flushed with nitrogen and treated under stirring with an 8 N chromic acid solution added in a thin stream at 0 C.,

until the red color of chromium tr-ioxide persisted in the mixture. (The 8 N solutionof chromic acid was prepared by dissolving 26.7 g. of chromium trioxide in 23 cc. of concentrated sulfuric acid and diluting withwater to cc.) The reaction mixture was stirred for 5 minutes further, diluted with water and the product collected by filtration, washed with water and dried under vacuum. There was. thus obtained 12fi-carboxymethyl- A -pregnen-l7a-ol-3,20-dione.

Through a suspension of 500 mg. of the foregoing compound in 10 ml. of glacial acetic acid there waspassed a current of dry hydrogen chloride for 4 hours at 15 C., .the resulting solution was then poured into ice-water, the solid collected by filtration, washed with water, dried and evaporated to dryness. Chromatography of the residue'on neutral alumina, followed by recrystallization of the solid eluates from acetone-hexane gave the pure 12,3,17a-l86t011e of A -pregnen-17u-ol-3,20-dione-12-acetic acid.

Example VI A solution of 1.25 g. of 1Z-carbomethoxymethylene- A -pregnadien-3,8-ol-20-one acetate in 50 cc. of ethyl acetate was hydrogenated at room'temperature and atmospheric pressure using 200 mg. of prereduced 10% palladium on charcoal catalyst, until 1 molar equivalent of hydrogen was absorbed; the catalyst was filtered and the filtrate evaporated to a small volume. Addition of methanol gave the crystalline 12-carbomethoxymethylene-A -pregnen-3fi-ol-ZO-one acetate.

A solution of 1 g. of the above compound in 25 cc. of 2.5% methanolic solution of perchloric acid was kept at room temperature for 18 hours, it was then diluted with water, the formed precipitate collected by filtration, washed with water, dried and recrystallized from acetonehexane, thus giving 12-carboxymethylene-A -pregnen-3pol-20-one.

The above compound was oxidized with 8 N chrornic acid in acetone solution, in accordance with the method of the preceding example to produce 12-carboxymethylene-A -pregnene-3,2O-dione.

The crude product was dissolved in 40 cc. of methanol and treated at room temperature with a solution of 0.1 g. of oxalic acid in 1 cc. of Water. The mixture was kept standing for 3 hours, then diluted with Water and the product was collected by filtration, washed with water to neutral and dried. There was thus obtained IZ-carboxyrnethylene-A pregnene-3,20-dione.

Upon esterification of the above compound with diazomethane in methylene chloride solution there was obtained the corresponding l2-carbomethoxymethylene-M- pregnene-3,20-dione.

Example VII 5 g. of the mixture of hydroxyesters obtained as described in Example I was heated with 20 cc. of acetic anhydride in a sealed tube at C. for 5 hours; cooled, and the contents of the tube transferred to an Erlenmeyer; the excess of anhydride was hydrolyzed by the addition of 8 cc. of water, the mixture stirred for 30 minutes, extractedwith ethyl acetate, washed with water to neutral, dried and evaporated to dryness under vacuo. The resulting oil was dissolved in 50 cc. of 5% methanolic potassium hydroxide solution and the mixture refluxed for 1 hour, cooled, acidified with hydrochloric acid and extracted several times with ethyl acetate, the organic solution was washed with water, dried and evaporated to dryness. There was thus obtained the 12-carboxyrnethylene derivative of A -22a-spirosten-3B-ol.

The above compound was dissolved in 30 cc. of anhydrous ether and 30 cc. of anhydrous dioxane and reduced with lithium in liquid ammonia, in accordance with the method of Example III. After evaporation of the ammonia and dilution with water, the mixture was acidified with hydrochloric acid and extracted with ethyl acetate, the organic solution was washed with water, dried and evaporated to dryness, thus affording 12,8-carboxymethyl-A -22a-spirosten-3B-ol.

2 g. of the above compound was dissolved in 20 cc. of methanol, 0.72 cc. of sulfuric acid was added and the mixture refluxed for 3 hours, diluted with water, the prod- -uct extracted with ethyl acetate, and the combined extractions washed with 5% sodium bicarbonate solution and water to neutral, dried and evaporated to dryness oxide. The mixture was kept at room temperature overnight, diluted with water, acidified with hydrochloric acid and extracted several times with ether, the extract was washed with water, dried and evaporated to dryness, thus producing the 125 carboxymetbyl 16u,17u oxido A pregnen-3 fi-ol-Ztl-one.

Further treatment with hydrobromic acid in acetic acid, followed by hydrogenation of the resulting bromohydrin, in accordance with the method of Example IV, gave 12fl-carboxymethyl-A -pregnene-3fi,17u-diol-20-onc.

Oxidation of the above compound with 8 N chromic acid in acetone solution, by following the method described in Example V, gave 1ZB-carboxymethyl-M-pregnen-17a-ol-3,20-dione identical with the comoound ohtained in such example.

Example VIII 1 g. of the latter compound was treated with oxalic acid in methanol solution, by following the method of Example VI, thus producing l2 8-carboxymethyl-A pregnen-17a-cl-3,20-dione. Upon treatment with dry hydrogen chloride in acetic acid, in accordance with the method of Example V there was obtained the 12,13,170-

lactone of A -pregnen-l7a-ol-3,20-dione-12-acctic acid, identical with that obtained in such example.

Example IX A solution of 1 g. of 12B-carboxyrnethyl-A -pregnen- 17u-ol-3,20-dione in 10 cc. of methylene chloride was treated with an ethereal solution of diazomethane to give 12B-carbomethoxymethyl A pregnen 17a ol-3,20-

' dione.

The above compound was dissolved in 50 cent benzene and treated with 2 cc. of acetic anhydride and 1 g. of ptoluenesulfonic acid; the mixture was kept at room temperature for 48 hours, washed well with water, sodium carbonate solution and water to neutral, dried over anhydrous sodium sulfate and evaporated to dryness; by chromatography of the residue on neutral alumina and recrystallization of the solid eluates from acetone-hexane,

there was obtained lZB-carbomethoxymethyl-M-pregnen 17lZ-O1-3,20-di0116 acetate.

In a similar manner, but using propionic, caproic or cyclopentylpropionic anhydride there were produced the corresponding 17-esters of 12 8-carbomethoxymethyl-A pregnan-17ot-ol-3,20-dione.

I claim:

1. A compound ofthe following formula:

COOR

wherein R and R are selected from the group consisting of hydrogen and a lower alkyl group and R is selected from the group consisting of hydrogen and an acyl radical of up to 12 carbon atoms.

2. A -pregnen 17a ol-3,20-dione-12fi-acetic acid.

3. A compound of the following formula:

wherein R is selected from the group consisting of hydrogen and a lower alkyl group.

4. The 12,17a-1actone of A -pregnen-17a-ol-3,20-dionelZB-acetic acid.

5. A compound of the following formula:

COOH

wherein R is selected from the group consisting of hydrogen and a lower alkyl group.

6. A -pregnadien 3 8 ol-20-one-12/3-acetic acid.

7. 12-carboxymethylene A pregnadien-3 3-ol-20- one.

8. 12-carboxymethylene-M-pregnene-3,20-dione. 9. 1Zfi-hydroxyethyl-A -pregnene-3 B, l7a-diol-20-one.

No references cited. 

3. A COMPOUND OF THE FOLLOWING FORMULA: 